Drug-eluting stent including crystalline cilostazol

ABSTRACT

Provided is a stent comprising: a stent skeleton; and a deposition layer containing a plurality of layers deposited on the stent skeleton; each layer of the deposition layer comprising crystalline cilostazol, at least one of the plurality of layers comprising a bioabsorbable polymer.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. application Ser. No.17/597,449 filed Jan. 6, 2022, which is a National Stage ofInternational Application No. PCT/JP2020/026688 filed Jul. 8, 2020,claiming priority based on Japanese Patent Application No. 2019-127529filed Jul. 9, 2019, the entire contents of each of which areincorporated herein by reference in their respective entireties.

TECHNICAL FIELD

The present invention relates to a stent coated with cilostazol, and amethod of producing it, more specifically, to a stent having a pluralityof layers containing cilostazol, and a method of producing it.

BACKGROUND ART

In recent years, arteriosclerotic diseases such as myocardialinfarction, angina pectoris, apoplectic stroke, and peripheral vasculardiseases are increasing. As a secure therapeutic method forarteriosclerotic diseases, for example, percutaneous transluminalangioplasty (hereinafter simply referred to as “PTA”) such aspercutaneous transluminal coronary angioplasty in a coronary artery ofthe heart, wherein a stenotic portion or an occluded portion of a bloodvessel is surgically expanded, is widely used. In PTA, a treatmentmethod that is carried out for a stenotic portion or an occluded portionof a coronary artery is specifically called percutaneous transluminalcoronary angioplasty (hereinafter simply referred to as “PTCA”).

PTCA is a technique in which a thin tube having a balloon attached toits tip (balloon catheter), or a stent, is inserted from an artery of anarm or a femoral region into a stenotic portion of a heart coronaryartery, and the balloon at the tip is inflated to expand the stenoticblood vessel, to thereby recover the blood flow. By this, theintravascular space of the lesion site is expanded to increase the bloodthat flows through the intravascular space. PTCA is used not only fortreatment of arteriosclerotic diseases, but also for, for example,treatment of stenosis of a shunt blood vessel formed in an arm of ahemodialysis patient.

In general, a blood vessel portion subjected to PTCA is damaged due todetachment of endothelial cells, damaging of the elastic lamina, or thelike, and this causes the proliferation of the intima as a healingresponse of the blood vessel wall. As a result, about 30 to 40% of thecases with successful expansion of a stenotic lesion by PTCA suffer fromrestenosis.

The restenosis in humans is thought to be caused, mainly, by theinflammatory process found as adhesion and/or infiltration of monocyteswhich occurs one to three days after PTCA, and by the intimalthickening/formation process by smooth muscle cells, whose proliferationreaches the peak about 45 days later. In cases where the restenosisoccurs, PTCA needs to be carried out again. Therefore, establishment ofa method of prevention, and a method of treatment, of the restenosis isan urgent task.

In view of this, attempts have been intensively made in order to reducethe restenosis rate by using a drug-eluting medical device (stent) to beplaced in a lumen, the device having an anticancer drug, animmunosuppressive drug, an anti-inflammatory drug, or a proliferationinhibitor for smooth muscle cells supported on a surface of the stent orthe like, which deice allows topical release of the drug for aboutseveral days at the site of placement in the lumen.

The drugs applied to drug-eluting stents are commonly limus drugs thatcan act as an anticancer drug or an immunosuppressive drug. With theirstrong cytotoxicities, these drugs strongly suppress the proliferationof vascular smooth muscle cells, that is, the intimal thickening, whichis the main cause of the restenosis. However, since they also stronglysuppress regeneration of vascular endothelial cells, they may inducedelayed in-stent thrombosis, which is clinically very problematic.

Attempts to use drugs other than limus drugs have also been made.Examples of such drugs include cilostazol, which is noncytotoxic, butwhose preparation is assumed to be difficult because of its poorsolubility in water. For example, Patent Document 1 proposes adrug-eluting stent comprising a mixture containing a bioabsorbablepolymer having a molecular weight of 40,000 to 600,000 and cilostazol,wherein a surface of a stent body composed of a metal or a polymermaterial is coated with the mixture (see, for example, claims and[0015]). Patent Document 1 also discloses that the stent allows elutionof the drug during the period of occurrence of restenosis in theinflammatory process or the intimal thickening/formation process afterthe placement of the stent, wherein the drug acts on intravascular cellsto effectively suppress intimal thickening, so that restenosis afterplacement of a stent, which has hitherto frequently occurred, can beremarkably improved (see [0028]).

PRIOR ART DOCUMENT Patent Document

-   [Patent Document 1] WO 2016/067994

SUMMARY OF THE INVENTION Problems to be Solved by the Invention

The stent disclosed in Patent Document 1 is used for, for example, arelatively thick artery such as a heart coronary artery, and required toact on the inflammatory process found as adhesion and/or infiltration ofmonocytes which occurs one to three days after PTCA. Thus, the stentaccording to Patent Document 1 is required to produce the effect byallowing release of cilostazol for several days after the placement.

On the other hand, in recent years, peripheral artery diseases (PADS)caused by infarction or the like of peripheral arteries, which arethinner, are attracting attention. For example, there are diseases inwhich arteriosclerosis occurs in leg blood vessels to cause thinning orclogging of the blood vessels, leading to insufficiency of blood flow tothe legs. They cause symptoms such as numbness, pain, and cold feelingduring walking. As the disease progresses, the patient becomes unable towalk (intermittent claudication), or feels pain in the legs even atrest. Further progression of the disease may lead to ulceration ornecrosis of the legs, and, in severe cases, surgery is required for thelegs.

Even in cases where PAD merely exhibits symptoms of the limbs,arteriosclerosis may affect not only the limbs, but also blood vesselsin the whole body. In cases where PAD is left untreated, it may alsocause myocardial infarction, angina pectoris, cerebral infarction,and/or the like. Various therapeutic methods such as pharmacotherapy,physical therapy, and surgery are available for PAD depending on theprogression of the disease state and the therapeutic goal. By realizinga drug-placing stent for peripheral arteries, a novel therapeutic methodfor minimally invasive treatment of PAD can be provided.

As a result of studies, the present inventors considered that a stentthat allows the presence of an effective component in an arterial vesselin a diseased state, for a longer period (for example, 6 to 12 months)than a drug-placing stent for a heart coronary artery (for example,Patent Document 1) is required.

An object of the present invention is to provide a drug-placing stentthat allows the presence of an effective component in a blood vessel ina diseased state, for a longer period (for example, 6 to 12 months).Such a drug-placing stent can be suitably used for treatment of aperipheral blood vessel (for example, peripheral arterial blood vessel),and can provide a minimally invasive therapeutic method.

Means for Solving Problems

As a result of intensive study, the present inventors discovered that adrug-placing stent comprising: a stent skeleton; and a deposition layercontaining a plurality of layers deposited on the stent skeleton; eachlayer of the deposition layer comprising crystalline cilostazol, atleast one of the plurality of layers comprising a bioabsorbable polymer,wherein elution of not more than 5% by mass of the crystallinecilostazol occurs by 24 hours in a test on the elution rate in vitro,can be obtained. The present inventors also discovered that such adrug-placing stent can be suitably used for peripheral blood vessels,thereby completing the present invention.

The present description includes the following embodiments.

1. A stent comprising:

-   -   a stent skeleton; and    -   a deposition layer containing a plurality of layers deposited on        the stent skeleton;    -   each layer of the deposition layer comprising crystalline        cilostazol (CLZ),    -   at least one of the plurality of layers comprising a        bioabsorbable polymer,    -   wherein elution of not more than 5% by mass of the crystalline        cilostazol occurs by 24 hours after the stent is brought into        contact in vitro with an elution medium, at 37° C., of a        phosphate-buffered sodium chloride solution containing 0.25% by        mass of sodium lauryl sulfate.

2. A stent comprising:

-   -   a stent skeleton; and    -   a deposition layer containing a plurality of layers deposited on        the stent skeleton;    -   each layer of the deposition layer comprising crystalline        cilostazol (CLZ),    -   at least one of the plurality of layers comprising a        bioabsorbable polymer,    -   wherein elution of not more than 20% by mass of the crystalline        cilostazol occurs by 15 days after the stent is brought into        contact in vitro at 37° C. with an elution medium of a        phosphate-buffered sodium chloride solution containing 0.25% by        mass of sodium lauryl sulfate.

3. The stent according to 1 or 2, wherein the deposition layer has atleast two layers, and wherein the content of cilostazol in a firstlayer, which is closer to the stent, is higher than the content ofcilostazol in a second layer, which is more distant from the stent.

4. The stent according to any one of 1 to 3, wherein

-   -   the deposition layer has at least two layers;    -   the content of cilostazol in the first layer, which is closer to        the stent, is higher than the content of cilostazol in the        second layer, which is more distant from the stent; and    -   both layers comprise a bioabsorbable polymer.

5. The stent according to any one of 1 to 4, wherein the bioabsorbablepolymer comprises not less than 90% by mass of polylactic acid.

6. The stent according to any one of 1 to 5, wherein the bioabsorbablepolymer comprises L-lactide and DL-lactide at a mass ratio of 6:4 to8:2, and has a viscosity of 1.8 to 4.5 dL/g.

7. The stent according to any one of 1 to 5, wherein the bioabsorbablepolymer comprises not less than 90% by mass of L-lactide, and has aviscosity of 0.6 to 1.4 dL/g.

8. The stent according to any one of 1 to 7, to be used for a peripheralblood vessel.

9. A stent comprising:

-   -   a stent skeleton;    -   a first layer deposited on the stent skeleton; and    -   a second layer deposited on the first layer;    -   the first layer and the second layer each comprising cilostazol        and a bioabsorbable polymer,    -   the bioabsorbable polymer comprising L-lactide and DL-lactide at        a mass ratio of 6:4 to 8:2, and having a viscosity of 1.8 to 4.5        dL/g,    -   the first layer comprising 470±47 μg of cilostazol and 313±31 μg        of the bioabsorbable polymer, the second layer comprising 30±3        μg of cilostazol and 270±27 μg of the bioabsorbable polymer.

Effects of the Invention

A drug-placing stent of an embodiment of the present invention iscapable of allowing elution of not more than 5% by mass of thecrystalline cilostazol by 24 hours in a test on the elution rate invitro. Thus, the drug-placing stent of the embodiment of the presentinvention is capable of allowing elution of crystalline cilostazol for alonger period, and can be more suitably used for peripheral bloodvessels.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 schematically illustrates the whole shape (a) of a stent of anembodiment of the present invention, and a cross-section (b) of thestent along A-A.

FIG. 2 schematically illustrates an embodiment of coating of a stentwith a coating agent using an ultrasonic atomizer.

EMBODIMENT FOR CARRYING OUT THE INVENTION

A stent of one embodiment of the present invention comprises:

-   -   a stent skeleton; and    -   a deposition layer containing a plurality of layers deposited on        the stent skeleton;    -   each layer of the deposition layer comprising crystalline        cilostazol,    -   at least one of the plurality of layers comprising a        bioabsorbable polymer, wherein    -   elution of not more than 5% by mass of the crystalline        cilostazol occurs by 24 hours after the stent is brought into        contact in vitro with an elution medium, at 37° C., of a        phosphate-buffered sodium chloride solution containing by mass        of sodium lauryl sulfate.

A stent of another embodiment of the present invention comprises:

-   -   a stent skeleton; and    -   a deposition layer containing a plurality of layers deposited on        the stent skeleton;    -   each layer of the deposition layer comprising crystalline        cilostazol,    -   at least one of the plurality of layers comprising a        bioabsorbable polymer,    -   wherein elution of not more than 20% by mass of the crystalline        cilostazol occurs by 15 days after the stent is brought into        contact in vitro at 37° C. with an elution medium of a        phosphate-buffered sodium chloride solution containing 0.25% by        mass of sodium lauryl sulfate.

A stent of still another embodiment of the present invention comprises:

-   -   a stent skeleton;    -   a first layer deposited on the stent skeleton; and    -   a second layer deposited on the first layer;    -   the first layer and the second layer each comprising cilostazol        and a bioabsorbable polymer,    -   the bioabsorbable polymer comprising L-lactide and DL-lactide at        a mass ratio of 6:4 to 8:2, and having a viscosity of 1.8 to 4.5        dL/g,    -   the first layer comprising 470±47 μg of cilostazol and 313±31 μg        of the bioabsorbable polymer,    -   the second layer comprising 30±3 μg of cilostazol and 270±27 μg        of the bioabsorbable polymer.

A stent of an embodiment of the present invention comprises: a stentskeleton; and a deposition layer containing a plurality of layersdeposited on the stent skeleton.

In an embodiment of the present invention, “stent skeleton” means askeleton forming a stent, and is usually not limited as long as, forexample, it is formed as a coarse-mesh cylinder using a metal or apolymer material, and as long as a desired stent of the presentinvention can be obtained therewith.

Examples of the stent skeleton made of a metal include stent skeletonsmade of nickel, cobalt, chromium, titanium, or an appropriate alloy suchas stainless steel. The stent skeleton is preferably a stent skeletonmade of a metal comprising a cobalt-chromium alloy as a major component.

A stent of an embodiment of the present invention comprises a depositionlayer containing a plurality of layers deposited on the stent skeleton.Each layer of the deposition layer comprises crystalline cilostazol, andat least one of the plurality of layers comprises a bioabsorbablepolymer.

In the present description, “cilostazol” has the chemical name6-[4-(1-cyclohexyl-1H-tetrazol-5-yl)butoxy]-3,4-dihydrocarbostyryl.Cilostazol has platelet aggregation inhibition action, phosphodiesterase(PDE) inhibition action, antiulcer action, hypotensive action, andanti-inflammatory action, and is known to be useful as an antithromboticdrug, a cerebral circulation improver, an antiphlogistic, an antiulcerdrug, an antihypertensive drug, an antiasthmatic drug, aphosphodiesterase inhibitor, and the like. The cilostazol is not limitedas long as a desired stent of the present invention can be obtainedtherewith. The cilostazol also includes pharmaceutically acceptablesalts thereof.

The cilostazol is preferably crystalline. The cilostazol preferably hasa crystal structure rather than having no crystal structure (rather thanbeing amorphous), from the viewpoint of reducing the elution rate.

In an embodiment of the present invention, the bioabsorbable polymer isnot limited as long as a desired stent of the present invention can beobtained therewith. Examples of the bioabsorbable polymer includepolylactic acid including lactide, and its molecular weight (Mw: weightaverage molecular weight) may be 40,000 to 700,000. Its viscosity may be0.4 to 5.0 dL/g, or may be 0.4 to 4.2 dL/g. Further, the bioabsorbablepolymer may comprise DL-lactide, L-lactide, or the like, and maycomprise glycolide, caprolactone, or the like. Specific examples of thebioabsorbable polymer include: polymers containing DL-lactide having amolecular weight of 10,000 to 1,000,000; polymers containing L-lactideand DL-lactide at a mass ratio of 6:4 to 8:2, and having a molecularweight of 300,000 to 650,000 or a viscosity of 1.8 to 4.5 dL/g; andpolymers having a molecular weight of 50,000 to 150,000 or containingnot less than 90% by mass L-lactide, and having a viscosity of 0.6 to1.4 dL/g (including poly(L-lactic acid) containing 100% by massL-lactide). As the bioabsorbable polymer, a commercially availableproduct may be used. Examples of the commercially available productinclude LR704S (trade name), L206S (trade name), and LR706S (tradename). The bioabsorbable polymers may be used either individually or incombination.

The bioabsorbable polymer preferably comprises not less than 90% bymass, more preferably comprises not less than 93% by mass, still morepreferably comprises 96% by mass polylactic acid. In cases where thebioabsorbable polymer comprises not less than 90% by mass polylacticacid, the polymer is capable of sustained release, which is anadvantageous effect.

The deposition layer has at least two layers, wherein the content ofcilostazol in a first layer, which is closer to the stent skeleton, ispreferably higher than the content of cilostazol in a second layer,which is more distant from the stent skeleton. In cases where the layercloser to the stent skeleton has a higher content of cilostazol, thelayer allows sustained release for a long period, which is anadvantageous effect.

The content of cilostazol in the first layer is preferably 300 to 750μg, more preferably 350 to 550 μg, still more preferably 440 to 480 μg.Further, the first layer preferably comprises 470±47 μg cilostazol.

The content of cilostazol in the second layer is preferably 0 to 100 μg,more preferably 10 to 80 μg, still more preferably 20 to 60 μg. Further,the second layer preferably comprises 30±3 μg cilostazol.

The content of cilostazol in the first layer is preferably 40 to 100% bymass, more preferably 50 to 70% by mass, still more preferably 55 to 65%by mass.

The content of cilostazol in the second layer is preferably 0 to 50% bymass, more preferably 2 to 20% by mass, still more preferably 5 to 10%by mass.

The deposition layer has at least two layers, wherein the content ofcilostazol in a first layer, which is closer to the stent, is preferablyhigher than the content of cilostazol in a second layer, which is moredistant from the stent, and wherein both layers preferably comprise abioabsorbable polymer.

The content of the bioabsorbable polymer in the first layer ispreferably 0 to 500 μg, more preferably 250 to 350 μg, still morepreferably 300 to 320 μg. Further, the first layer preferably comprises313±31 μg of the bioabsorbable polymer.

The content of the bioabsorbable polymer in the second layer ispreferably 180 to 540 μg, more preferably 200 to 300 μg, still morepreferably 260 to 285 μg. Further, the second layer preferably comprises270±27 μg of the bioabsorbable polymer.

The content of the bioabsorbable polymer in the first layer ispreferably 0 to 60% by mass, more preferably 25 to 50% by mass, stillmore preferably 35 to 45% by mass.

The content of the bioabsorbable polymer in the second layer ispreferably not less than 70% by mass, more preferably not less than 80%by mass, still more preferably not less than 90% by mass.

In a stent of an embodiment of the present invention, elution of notmore than 3% by mass of the crystalline cilostazol occurs by 24 hoursafter the stent is brought into contact in vitro with an elution medium,at 37° C., of a phosphate-buffered sodium chloride solution containingby mass sodium lauryl sulfate.

In a stent of an embodiment of the present invention, elution of notmore than 20% by mass of the crystalline cilostazol preferably occurs by15 days after the stent is brought into contact; elution of not morethan 7% by mass of the crystalline cilostazol preferably occurs by 8days after the stent is brought into contact; and elution of not morethan 5% by mass, more preferably not more than 3% by mass, of thecrystalline cilostazol preferably occurs by 1 day after the stent isbrought into contact; in vitro at 37° C. with an elution medium of aphosphate-buffered sodium chloride solution containing 0.25% by masssodium lauryl sulfate.

In a stent of an embodiment of the present invention, elution of, forexample, not less than 1.0% by mass of the crystalline cilostazol mayoccur, or not less than 0.1% by mass of the crystalline cilostazol mayoccur, by 15 days after the stent is brought into contact; elution of,for example, not less than 0.5% by mass of the crystalline cilostazolmay occur, or not less than 0.05% by mass of the crystalline cilostazolmay occur, by 8 days after the stent is brought into contact; elutionof, for example, not less than 0.1% by mass of the crystallinecilostazol may occur, or not less than 0.01% by mass of the crystallinecilostazol may occur, by 1 day after the stent is brought into contact;in vitro at 37° C. with an elution medium of a phosphate-buffered sodiumchloride solution containing 0.25% by mass sodium lauryl sulfate.

In cases where elution of not more than the above-described ratio of thecrystalline cilostazol occurs by the above-described time after thestent is brought into contact in vitro with an elution medium, at 37°C., of a phosphate-buffered sodium chloride solution containing 0.25% bymass sodium lauryl sulfate, the cilostazol can be released for more thanthree months, so that the crystalline cilostazol can be allowed to bepresent in the living body for a very long time.

Thus, a stent of an embodiment of the present invention can be suitablyused, for example, for a peripheral blood vessel, preferably for aperipheral arterial blood vessel.

Further, a stent of an embodiment of the present invention can also beused for, for example, relatively thick arteries for which stents haveconventionally been used, such as heart coronary arteries and lowerextremity arteries.

As long as a desired stent of the present invention can be obtained, themethod of producing a stent of an embodiment of the present invention isnot limited.

A stent of an embodiment of the present invention can be produced using,for example, a production method comprising: (i) providing a stentskeleton; (ii) providing a mixture containing cilostazol; (iii) coatingthe stent skeleton with the mixture; and repeating (ii) and (iii)(wherein the content of cilostazol, and/or the like is/are adjusted).

The mixture containing cilostazol may contain the bioabsorbable polymerdescribed above, in addition to the cilostazol. The mixture may alsocontain a solvent such as an additive. Since cilostazol is poorlysoluble, the bioabsorbable polymer is required to prevent detachment ofthe coating, and to maintain high strength.

The mixing mass ratio between the cilostazol and the bioabsorbablepolymer such as polylactic acid is preferably 1:0.5 to 1:1.5. In caseswhere the ratio is within this range, a better intimal-thickening effectcan be obtained. In cases where the mixing mass ratio is 1:1.1 to 1:1.5,the coating strength and the sustained-release effect can be furtherincreased.

In an embodiment of the present invention, the method of coating thestent skeleton with the mixture of cilostazol and the bioabsorbablepolymer is not limited as long as a desired stent of the presentinvention can be obtained. For example, a simple spraying method, adipping method, an electrodeposition method, an ultrasonic spray method,or the like that has been conventionally used may be employed. From theviewpoint of coating, it is preferred to use an ultrasonic spray method.

The embodiments described above may be combined as appropriate when itis possible.

Embodiments of the present invention are described below in detail withreference to drawings.

FIG. 1(a) schematically illustrates a drug-eluting stent 1 of oneembodiment of the present invention. The drug-eluting stent 1 has acylindrical shape having a longitudinal axis, and a lumen is formedtherein. In the drug-eluting stent 1, the cylindrical shape has acoarse-mesh-like side such that it is laterally expandable. Usually, thenet may be formed with a member 2 (a wire of a metal or a polymermaterial) forming the stent skeleton. The drug-eluting stent 1 isusually inserted in an unexpanded form into the body. It is expanded ata treatment site in a blood vessel, and placed in the blood vessel. Theexpansion may be achieved by a balloon catheter in the blood vessel.FIG. 1(a) schematically illustrates the net. As long as a desired stentof the present invention can be obtained, the pattern of the net is notlimited.

FIG. 1(b) schematically illustrates a cross-section (A-A cross-section)of a wire forming the stent skeleton in FIG. 1(a).

In a drug-eluting stent 1 of one embodiment of the present invention, adeposition layer 3 is formed on the stent skeleton member 2. The stentskeleton member 2 may be prepared using an arbitrary method. Forexample, it may be prepared by laser, electric discharge milling,chemical etching, or another means, from a hollow or formedstainless-steel tube. The stent skeleton member 2 may be formed withnickel, cobalt, chromium, titanium, or an appropriate alloy of stainlesssteel etc.

The deposition layer 3 is formed with at least two layers. In FIG. 1(b),the plurality of layers are not illustrated.

FIG. 2 schematically illustrates an ultrasonic spray coating device 4for forming the deposition layer 3 by application on the stent skeletonmember 2. Before the coating step, first, the surface of the stentskeleton member 2 is preferably subjected to plasma treatment using aplasma treatment apparatus (not shown). Thereafter, the stent skeletonmember 2 is mounted on a mandrel, followed by attachment to theultrasonic spray coating device 4. In the ultrasonic spray coatingdevice 4, a coating liquid is transferred by a syringe pump through apipe 6, and then atomized and sprayed by an ultrasonic spray nozzle 5.During the spray, the stent skeleton member 2 is linearly moved whilerotating the member under the ultrasonic nozzle to allow deposition ofthe deposition layer 3 on the stent skeleton member 2. Thereafter, whilerotating and linearly moving the stent skeleton member 2, it is driedunder nitrogen gas flow, followed by further drying it in a desiccatorunder reduced pressure. By this, a drug-eluting stent 1 can be prepared.Depending on the number of layers to be included in the deposition layer3, different coating liquids are used to coat the stent skeleton membera plurality of times to form the deposition layer 3.

Each coating liquid is provided using a mixture prepared by dissolvingcilostazol and a bioabsorbable polymer in a solvent at a ratio inaccordance with the deposition layer 3 to be formed. Since thedeposition layer 3 includes a plurality of layers, a plurality ofcoating liquids need to be provided. As the coating solvent, a volatilesolvent having a low boiling point may be used for enabling simpleremoval after coating. Examples of the volatile solvent includemethanol, ethanol, trifluoroethanol, hexafluoroisopropanol, isoamylalcohol, methyl acetate, ethyl acetate, acetone, methyl ethyl ketone,methylene chloride, chloroform, and dichloroethane; and mixed solventscontaining at least two of these.

EXAMPLES

The present invention is described below more concretely in more detailby way of Examples and Comparative Examples. However, these Examples aremerely embodiments of the present invention, and the present inventionis not limited by these examples at all.

The polymers used in the present Examples are shown below in Table 1.

TABLE 1 Molecular weight Composition (Mw: weight L: L-lactide averageDL: DL-lactide molecular G: glycoside Polymer Viscosity (dL/g) weight)C: caprolactine (a) L2065  0.8-1.2 102,000 L = 100 (b) LR704S 2.0-2.8 >350,000 L:DL = 70:30 (c) LR708S  3.3-4.2 >630,000 L:DL = 70:30(d) RG755S 0.50-0.70 63,000 DL:G = 75:26 (e) RG868S  1.3-1.7 220,000DL:G = 88:15 (f) LC703S  1.3-1.8 257,000 L:C = 70:30

The viscosity of each polymer described above means the limitingviscosity [ηη] (dL/g), which was measured by the capillary viscometermethod. A sample solution with a concentration of C (g/dL) was provided,and the measured values of the flow time of the sample solution (t) andthe flow time of the solvent (t0) were used to calculate the limitingviscosity according to the following equation.

$\lbrack\eta\rbrack = {\lim\limits_{c\rightarrow 0}\frac{\ln\frac{t}{t_{0}}}{c}}$

An Ubbelohde viscometer was used as the apparatus, and chloroform (25°C.) was used as the solvent.

Further, the weight average molecular weight (Mw) was determined usingthe Mark-Kuhn-Houwink equation (Mark-Howink-Sakurada equation).

Example 1

A cobalt-chromium alloy was used as the stent skeleton member 2.Cilostazol (CLZ) was dissolved in methylene chloride to provide asolution. By ultrasonic spray coating, this solution was applied to thebase material cobalt-chromium alloy, to form a first layer composed of440 μg of cilostazol.

Subsequently, cilostazol and polymer (a) were mixed together at 1:9(mass ratio), and dissolved in methylene chloride, to provide asolution. By ultrasonic spray coating, this solution was applied to thefirst layer, to form a second layer composed of 540 μg of polymer (a)and 60 μg of cilostazol, to obtain a stent of Example 1.

Examples 2 and 3

Stents of Examples 2 and 3 were produced using the same method as themethod described in Example 1 except that polymer (b) or polymer (c) wasused instead of polymer (a).

Comparative Examples 1 to 3

Stents of Comparative Examples 1 to 3 were produced using the samemethod as the method described in Example 1 except that polymers (d) to(f) were used instead of polymer (a).

Example 4

A cobalt-chromium alloy was used as the stent skeleton member 2.Cilostazol and polymer (b) were mixed together at a ratio of 3:2, anddissolved in methylene chloride, to provide a solution. By ultrasonicspray coating, this solution was applied to the cobalt-chromium alloy ofthe stent skeleton member, to form a first layer composed of 313 μg ofpolymer (b) and 470 μg of cilostazol.

Subsequently, cilostazol and polymer (b) were mixed together at a ratioof 1:9 (mass ratio), and dissolved in methylene chloride, to provide asolution. By ultrasonic spray coating, this solution was applied to thefirst layer, to form a second layer composed of 270 μg of polymer (b)and 30 μg of cilostazol, to obtain a stent of Example 4.

Example 5

A cobalt-chromium alloy was used as the stent skeleton member 2.Cilostazol and polymer (b) were mixed together at a ratio of 3:2, anddissolved in methylene chloride, to provide a solution. By ultrasonicspray coating, this solution was applied to the base materialcobalt-chromium alloy, to form a first layer composed of 323 μg ofpolymer (b) and 485 μg of cilostazol.

Subsequently, cilostazol and polymer (b) were mixed together at a ratioof 1:19 (mass ratio), and dissolved in methylene chloride, to provide asolution. By ultrasonic spray coating, this solution was applied to thefirst layer, to form a second layer composed of 285 μg of polymer (b)and 15 μg of cilostazol, to obtain a stent of Example 5.

Example 6

A cobalt-chromium alloy was used as the stent skeleton member 2.Cilostazol and polymer (b) were mixed together at a ratio of 3:2, anddissolved in methylene chloride, to provide a solution. By ultrasonicspray coating, this solution was applied to the base materialcobalt-chromium alloy, to form a first layer composed of 313 μg ofpolymer (b) and 470 μg of cilostazol.

Subsequently, cilostazol and polymer (c) were mixed together at a ratioof 1:9 (mass ratio), and dissolved in methylene chloride, to provide asolution. By ultrasonic spray coating, this solution was applied to thefirst layer, to form a second layer composed of 270 μg of polymer (c)and 30 μg of cilostazol, to obtain a stent of Example 6.

Example 7

A cobalt-chromium alloy was used as the stent skeleton member 2.Cilostazol and polymer (b) were mixed together at a ratio of 3:2, anddissolved in methylene chloride, to provide a solution. By ultrasonicspray coating, this solution was applied to the base materialcobalt-chromium alloy, to form a first layer composed of 490 μg ofpolymer (b) and 735 μg of cilostazol.

Subsequently, cilostazol and polymer (b) were mixed together at a ratioof 1:19 (mass ratio), and dissolved in methylene chloride, to provide asolution. By ultrasonic spray coating, this solution was applied to thefirst layer, to form a second layer composed of 285 μg of polymer (b)and 15 μg of cilostazol, to obtain a stent of Example 7.

Example 8

Using the same method as the method described in Example 4 except that afirst layer composed of 180 μg of polymer (b) and 270 μg of cilostazolwas formed, a stent of Example 8 was obtained.

Example 9

Using the same method as the method described in Example 4 except that afirst layer composed of 247 μg of polymer (b) and 370 μg of cilostazolwas formed, a stent of Example 9 was obtained.

Cilostazol Elution Test (In Vitro) 1. Method of Cilostazol Elution Test

Using an elution tester 400-DS (Apparatus 7), and using 10 mL of aphosphate-buffered sodium chloride solution containing 0.25% by masssodium lauryl sulfate as a test liquid, a test was carried out at anelution test liquid temperature of 37° C. with Dip Speed 10.

Sampling was carried out 0.5, 1, 3, 6, 9, 12, 18, and 24 hours later.The whole amount of the test liquid was replaced at each collectiontime.

2. Measurement of Elution Rate of Cilostazol (HPLC Measurement)

Under the following conditions, HPLC measurement was carried out for 10μL of each sample solution or a standard solution. From the peak areavalues At and As of cilostazol, the elution rate was calculated.

The elution rate was calculated using the following equation.

$\left\lbrack {Q{{hr}(\%)}} \right\rbrack = {M_{S} \times \frac{{\sum}_{i = P}^{Q}A_{T}i}{A_{S}} \times \frac{1}{C} \times A}$Q: Elution rate (% by mass) for the indicated amount of cilostazol athour Q M_(S): Weighed amount of standard sample of cilostazol (mg)A_(T): Peak area of cilostazol sampled at hour i A_(S): Peak area ofstandard solution C: Indicated amount of cilostazol per stent (≈loadedamount of cilostazol) A: Constant (0.5) Σ_(i=P) ^(Q)A_(T) ^(i): Sum ofthe peak area of cilostazol from hour p to hour Q As the indicatedamount of cilostazol per stent (C), the amount of loaded cilostazolcalculated from the coating mass was used.

Measurement Conditions

-   -   Detector: ultraviolet absorptiometer (measurement wavelength:        254 nm)    -   Column: stainless-steel tube of 4.6 mm (inner diameter)×150 mm        (length), packed with 5-μm octadecylsilyl-modified silica gel        for liquid chromatography    -   Column temperature: constant temperature near 25° C.    -   Mobile phase: water/acetonitrile/methanol mixed liquid (10:7:3,        v/v/v)    -   Flow rate: adjusted such that the retention time of cilostazol        was about 9 minutes        Cilostazol Concentration in Arterial Tissue after Implantation        in Rabbit Iliac Artery, and Residual Amount of Cilostazol on        Stent (In Vivo)

In a rabbit iliac artery, each of the stents of Examples and ComparativeExamples was implanted. The implantation was carried out as follows.

First, the neck of a rabbit is incised to expose the right carotidartery, and an introducer is placed therein. A balloon catheter guidewire is inserted from the introducer, and moved to the distal portion ofthe treatment site of the iliac artery under X-ray fluoroscopy.Thereafter, an angiographic catheter is inserted along the guide wire,and angiography is carried out for the treatment site of the iliacartery. After the completion of the angiography of the treatment site, asample balloon catheter is inserted, along the balloon catheter guidewire, to the treatment site under X-ray fluoroscopy. After confirmingthat the sample stent (standard diameter: 2.75-mm stent diameter at aninflation pressure of 9 atm) is placed in the treatment site of theiliac artery (expected blood vessel diameter, 2.5 mm), the balloon iskept in an inflated state at 14 atm (hyperinflation; expected stentdiameter, 3.0 mm; 20% by mass hyperinflation) using an indeflator for 20seconds each time. After confirming expansion of the stent, the balloonis deflated, and the indeflator is removed, followed by pulling out theballoon catheter along the balloon catheter guide wire. The left andright iliac arteries are treated by the same method.

Subsequently, the angiographic catheter is moved along the ballooncatheter guide wire to a position before the treatment site, andangiography is carried out using a diluted contrast agent. After theleft and right iliac arteries are treated by the same method, theangiographic catheter is pulled out. Finally, the blood vessel at thesheath insertion site is ligated, and the skin and the muscle layer aresutured. By the above process, the stent is placed in the iliac bloodvessel of the rabbit.

Ninety days after the implantation, the cilostazol concentration and theresidual stent amount in the arterial tissue at the implantation site ofeach stent were analyzed.

For pretreatment, the arterial tissue at the implantation site of eachstent was separated from the stent. From each sample separated, anorganic layer was obtained by liquid-liquid extraction, and then theorganic layer was dried to provide a sample. The obtained sample wassubjected to LC/MS/MS using the electrospray ionization method, toquantify cilostazol. The cilostazol concentration in the arterial tissue(μg cilostazol in 1 g of the tissue; μg/g tissue) and the residualamount of cilostazol (the residual ratio (%) of cilostazol on the stent)were calculated.

Cilostazol Concentration in Arterial Tissue after Implantation in PigIliac Artery, and Residual Amount of Cilostazol on Stent (In Vivo)

The cilostazol concentration in an arterial tissue after implantation ina pig iliac artery, and the residual amount of cilostazol, wereevaluated using the same method as the method carried out for therabbit, to analyze the cilostazol concentration in the arterial tissue(μg cilostazol in 1 g of the tissue; μg/g tissue) and the residualamount of cilostazol (the residual ratio (%) of cilostazol on the stent)in a pig.

TABLE 2 Example Comparative Example 1 2 3 1 2 3 First CLZ/polymerCLZ/polymer = CLZ/polymer = CLZ/polymer = CLZ/polymer = CLZ/polymer =CLZ/polymer = layer mass ratio 100/0 100/0 100/0 100/0 100/0 100/0 CLZ:μg CLZ: 440 CLZ: 440 CLZ: 440 CLZ: 440 CLZ: 440 CLZ: 440 Polymer: μgPolymer: 0 Polymer: 0 Polymer: 0 Polymer: 0 Polymer: 0 Polymer: 0 SecondCLZ/polymer CLZ/polymer CLZ/polymer CLZ/polymer CLZ/polymer CLZ/polymerCLZ/polymer layer mass ratio (a) = 1/9 (b) = 1/9 (c) = 1/9 (d) = 1/9 (e)= 1/9 (f) = 1/9 CLZ: μg CLZ: 60 CLZ: 60 CLZ: 60 CLZ: 60 CLZ: 60 CLZ: 60Polymer: μg Polymer (a): Polymer (b): Polymer (c): Polymer (d): Polymer(e): Polymer (f): 540 540 540 540 540 540 Total CLZ: μg 500 500 500 500500 500 CLZ 24 hours 1.7 3.2 1.1 51.7 17.3 39.6 elution rate (%)

TABLE 3 Example 4 5 6 7 First layer CLZ/polymer CLZ/polymer CLZ/polymerCLZ/polymer CLZ/polymer mass ratio (b) = 3/2 (b) = 3/2 (b) = 3/2 (b) =3/2 CLZ: μg CLZ: 470 CLZ: 485 CLZ: 470 CLZ: 735 Polymer: μg Polymer: 313Polymer: 323 Polymer: 313 Polymer: 490 Second layer CLZ/polymerCLZ/polymer CLZ/polymer CLZ/polymer CLZ/polymer mass ratio (b) = 1/9 (b)= 1/19 (c ) = 1/9 (b) = 1/19 CLZ: μg CLZ: 30 CLZ: 15 CLZ: 30 CLZ: 15Polymer: μg Polymer (b): Polymer (b): Polymer (c): Polymer (b): 270 285270 285 Total CLZ: μg 500 500 500 750 Rabbit CLZ 1 month later 100 96 88100 residual ratio 3 months later 77 89 99.8 91 (%) Pig CLZ 3 monthslater 87 Not measured Not measured Not measured concentration 6 monthslater 57 in tissue^(a) 9 months later 11 Pig CLZ 3 months later 70 Notmeasured Not measured Not measured residual ratio 6 months later 38 (%)9 months later 2 CLZ elution 24 hours 1.6 Not measured Not measured Notmeasured rate (%) 96 hours 4.3 15 days 13.2 ^(a)Cilostazol (μg) in 1 gof tissue. Unit: μg/g tissue.

TABLE 4 Example 8 9 First layer CLZ/polymer CLZ/polymer CLZ/polymer massratio (b) = 3/2 (b) = 3/2 CLZ: μg CLZ: 270 CLZ: 370 Polymer: μg Polymer:180 Polymer: 247 Second layer CLZ/polymer CLZ/polymer CLZ/polymer massratio (b) = 1/9 (b) = 1/9 CLZ: μg CLZ: 30 CLZ: 30 Polymer: μg Polymer(b): Polymer (b): 270 270 Total CLZ: μg 300 400 Pig CLZ  3 months later84 78 concentration  6 months later 46 36 in tissue^(a) Pig CLZ  3months later 66 60 residual ratio  6 months later 23 14 (%) CLZ elution24 hours 1.7 2.7 rate (%) 96 hours 3.8 5.6 ^(a)Cilostazol (μg) in 1 g oftissue. Unit: μg/g tissue.

Each of the stents of Examples 1 to 9 comprises a stent skeleton; and adeposition layer containing a plurality of layers deposited on the stentskeleton; each layer of the deposition layer comprising crystallinecilostazol, at least one of the plurality of layers comprising abioabsorbable polymer, wherein elution of not more than 5% by mass ofthe crystalline cilostazol occurs by 24 hours after the stent is broughtinto contact in vitro with an elution medium, at 37° C., of aphosphate-buffered sodium chloride solution containing 0.25% by mass ofsodium lauryl sulfate.

Further, each of the stents of Examples 1 to 9 comprises a stentskeleton; and a deposition layer containing a plurality of layersdeposited on the stent skeleton; each layer of the deposition layercomprising crystalline cilostazol, at least one of the plurality oflayers comprising a bioabsorbable polymer, wherein elution of not morethan 20% by mass of the crystalline cilostazol occurs by 15 days afterthe stent is brought into contact in vitro with an elution medium, at37° C., of a phosphate-buffered sodium chloride solution containing0.25% by mass of sodium lauryl sulfate.

Thus, the stents of Examples 1 to 9 are capable of sustained release ofcilostazol for more than three months.

Needless to say, the plurality of layers applicable are not limited totwo layers, and may be not less than three layers.

In contrast, in each of the stents of Comparative Examples 1 to 3,elution of more than 5% by mass of the crystalline cilostazol occurs by24 hours after the stent is brought into contact in vitro with anelution medium, at 37° C., of a phosphate-buffered sodium chloridesolution containing 0.25% by mass sodium lauryl sulfate.

Thus, the stents of Comparative Examples 1 to 3 are incapable ofsustained release of cilostazol for more than three months.

INDUSTRIAL APPLICABILITY

A drug-placing stent of an embodiment of the present invention iscapable of allowing elution of not more than 5% by mass of crystallinecilostazol by 24 hours in a test on the elution rate in vitro.Alternatively or additionally, a drug-placing stent of an embodiment ofthe present invention is capable of allowing elution of not more than20% by mass of crystalline cilostazol by 15 days in a test on theelution rate in vitro. Thus, the drug-placing stents of the embodimentsof the present invention are capable of allowing elution of crystallinecilostazol for a longer period, and can be more suitably used forperipheral blood vessels.

RELATED APPLICATIONS

The present application claims the priority under Article 4 of the ParisConvention based on the application of Application No. 2019-127529 filedin Japan on Jul. 9, 2019. The content of this priority application isincorporated in the present description by reference.

DESCRIPTION OF SYMBOLS

-   -   1. Stent    -   2. Stent skeleton member    -   3. Deposition layer    -   4. Ultrasonic spray coating device    -   5. Ultrasonic spray nozzle    -   6. Pipe

1. A stent comprising: a stent skeleton; and a deposition layercontaining a plurality of layers deposited on the stent skeleton; eachlayer of the deposition layer comprising crystalline cilostazol, atleast one of the plurality of layers comprising a bioabsorbable polymer,wherein the deposition layer has at least a first layer and a secondlayer; the content of cilostazol in the first layer, which is closer tothe stent, is higher than the content of cilostazol in the second layer,which is more distant from the stent; and both layers comprise abioabsorbable polymer, and wherein a content of the crystallinecilostazol in the first layer is 55% by mass to 100% by mass.
 2. Thestent according to claim 1, wherein the bioabsorbable polymer comprisesnot less than 90% by mass of polylactic acid.
 3. The stent according toclaim 1, wherein the bioabsorbable polymer comprises L-lactide andDL-lactide at a mass ratio of 6:4 to 8:2, and has a viscosity of 1.8 to4.5 dL/g.
 4. The stent according to claim 1, wherein the bioabsorbablepolymer comprises not less than 90% by mass of L-lactide, and has aviscosity of 0.6 to 1.4 dL/g.
 5. The stent according to claim 1, to beused for a peripheral blood vessel.
 6. The stent according to claim 1,wherein the bioabsorbable polymer comprises L-lactide and DL-lactide ata mass ratio of 6:4 to 8:2, and has a viscosity of 1.8 to 4.5 dL/g, thefirst layer comprises 470±47 μg of cilostazol and 313±31 μg of thebioabsorbable polymer, the second layer comprises 30±3 μg of cilostazoland 270±27 μg of the bioabsorbable polymer.
 7. The stent according toclaim 1, wherein an outer and exposed layer is bioabsorbable.
 8. Thestent according to claim 1, wherein the second layer is an outer andexposed layer.
 9. The stent according to claim 7, wherein the firstlayer comprises 300 μg to 750 μg of cilostazol drug, and greater than 0μg to 500 μg of the bioabsorbable polymer, and the second layercomprises 10 μg to 100 μg of cilostazol, and 180 μg to 540 μg of thebioabsorbable polymer.
 10. The stent according to claim 7, wherein thebioabsorbable polymer comprises L-lactide and DL-lactide at a mass ratioof 6:4 to 8:2, and has a viscosity of 1.8 to 4.5 dL/g, the first layercomprises 470±47 μg of cilostazol and 313±31 μg of the bioabsorbablepolymer, and the second layer comprises 30±3 μg of cilostazol and 270±27μg of the bioabsorbable polymer.